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International Journal of Analytical Chemistry
Volume 2015, Article ID 503139, 5 pages
Research Article

Development of HPLC Protocol and Simultaneous Quantification of Four Free Flavonoids from Dracocephalum heterophyllum Benth.

1Key Laboratory of Plant Resources and Chemistry in Arid Regions, Xinjiang Technical Institute of Physics and Chemistry, Chinese Academy of Sciences, Urumqi 830011, China
2Key Laboratory of Xinjiang Indigenous Medicinal Plants Resource Utilization, Xinjiang Technical Institute of Physics and Chemistry, Chinese Academy of Sciences, Urumqi 830011, China
3State Scientifically-Experimental and Production Organization, Academy of Sciences of the Republic of Tajikistan, 734063 Dushanbe, Tajikistan
4Department of Chemistry, Abdul Wali Khan University, Mardan 23200, Pakistan

Received 29 March 2015; Revised 18 April 2015; Accepted 20 April 2015

Academic Editor: Günther K. Bonn

Copyright © 2015 Sodik Rakhmonovich Numonov et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


Quantification of the four flavonoids, namely, luteolin, kaempferol, diosmetin, and chrysosplenetin, has been performed for the first time in 80% ethanolic extract of Dracocephalum heterophyllum B. through HPLC coupled to UV detector after optimization of extracting solvent and chromatographic conditions. Total flavonoids quantified were 0.324 mg/mL of the extract. HPLC analysis delivered contents of the luteolin, kaempferol, diosmetin, and chrysosplenetin as 0.08%, 0.14%, 0.28%, and 0.79% of the dried extract, respectively. LOD (%) values calculated were 0.04, 0.03, 0.03, and 0.08 and LOQ (%) values were 0.08, 0.12, 0.11, and 0.28 for luteolin, kaempferol, diosmetin, and chrysosplenetin, respectively. The recovery percentages for these flavonoids were within the acceptable range of 95% to 105%. Standard deviation and %RSD were calculated for each target analytes individually in extract for determining the reproducibility and accuracy of the method. In no case the %RSD was higher than 1 taking retention time as a factor while in the case of area under the curve maximum %RSD was noted in the case of diosmetin as 2.85. From our literature review regarding the plant species under study, it appears that these flavonoids have not been quantified before and are reported for the first time in this paper.